Beckman Coulter Cytomics FC 500 User manual

Cytomics FC 500
With CXP Software
Instructions For Use
PN 624923B (June 2004)
Beckman Coulter, Inc.
Fullerton, CA 92835

WARNINGS AND PRECAUTIONS
R E AD AL L PR O D U CT M ANU AL S AND CO NS U L T W IT H BE CK M AN CO U L T E R -T R AINE D PE R S O NNE L BE FO R E AT T E M PT ING
T O O PE R AT E INS T R U M E NT . D O NO T AT T E M PT T O PE R FO R M ANY PR O CE D U R E BE FO R E CAR E FU L L Y R E AD ING AL L
INS T R U CT IO NS . AL W AY S FO L L O W PR O D U CT L ABE L ING AND M ANU FACT U R E R ’S R E CO M M E ND AT IO NS . IF IN D O U BT AS
T O H O W T O PR O CE E D IN ANY S IT U AT IO N, CO NT ACT Y O U R BE CK M AN CO U L T E R R E PR E S E NT AT IV E .
BE CK M AN CO U L T E R , INC. U R G E S IT S CU S T O M E R S T O CO M PL Y W IT H AL L NAT IO NAL H E AL T H AND S AFE T Y
S T AND AR D S S U CH AS T H E U S E O F BAR R IE R PR O T E CT IO N. T H IS M AY INCL U D E , BU T IT IS NO T L IM IT E D T O ,
PR O T E CT IV E E Y E W E AR , G L O V E S , AND S U IT ABL E L ABO R AT O R Y AT T IR E W H E N O PE R AT ING O R M AINT AINING T H IS O R
ANY O T H E R AU T O M AT E D L ABO R AT O R Y ANAL Y Z E R .
HAZ ARDS AND OPERATIONAL PRECAUTIONS AND LIMITATIONS
W AR NING S , CAU T IO NS , and IM PO R T ANT S alert y ou as follow s :
WARNING - Can caus e injury .
CAUTION - Can caus e d amag e to th e ins trument.
IMPORTANT - Can caus e mis lead ing res ults .
WARNING R is k of op erator injury if:
r All d oors , covers and p anels are not clos ed and s ecured in p lace p rior to and d uring ins trument op eration.
r T h e integ rity of s afety interlocks and s ens ors is comp romis ed .
r Ins trument alarms and error mes s ag es are not acknow led g ed and acted up on.
r Y ou contact moving p arts .
r Y ou mis h and le b roken p arts .
r D oors , covers and p anels are not op ened , clos ed , removed and /or rep laced w ith care.
r Imp rop er tools are us ed for troub les h ooting .
T o avoid injury :
r K eep d oors , covers and p anels clos ed and s ecured in p lace w h ile th e ins trument is in us e.
r T ake full ad vantag e of th e s afety features of th e ins trument. D o not d efeat s afety interlocks and s ens ors .
r Acknow led g e and act up on ins trument alarms and error mes s ag es .
r K eep aw ay from moving p arts .
r R ep ort any b roken p arts to y our Beckman Coulter R ep res entative.
r O p en/remove and clos e/rep lace d oors , covers and p anels w ith care.
r U s e th e p rop er tools w h en troub les h ooting .
CAUTION S y s tem integ rity mig h t b e comp romis ed and op erational failures mig h t occur if:
r T h is eq uip ment is us ed in a manner oth er th an s p ecified . O p erate th e ins trument as ins tructed in th e Prod uct M anuals .
r Y ou introd uce s oftw are th at is not auth orized b y Beckman Coulter into y our comp uter. O nly op erate y our s y s tem’s
comp uter w ith s oftw are auth oriz ed b y Beckman Coulter.
r Y ou ins tall s oftw are th at is not an orig inal cop y rig h ted vers ion. O nly us e s oftw are th at is an orig inal cop y rig h ted
vers ion to p revent virus contamination.
IMPORTANT If y ou p urch as ed th is p rod uct from any one oth er th an Beckman Coulter or an auth orized Beckman Coulter
d is trib utor, and , if it is not p res ently und er a Beckman Coulter s ervice maintenance ag reement, Beckman Coulter cannot
g uarantee th at th e p rod uct is fitted w ith th e mos t current mand atory eng ineering revis ions or th at y ou w ill receive th e mos t
current information b ulletins concerning th e p rod uct. If y ou p urch as ed th is p rod uct from a th ird p arty and w ould like
furth er information concerning th is top ic, call y our Beckman Coulter R ep res entative.

iii
PN 624923B
REVISION STATUS
Issu e A, In itial Issu e, 8 /03
CX P S o ftw a re V e r sio n 1.0 . Initia l is s u e fo r c u s to m e r dis tr ib u tio n.
Issu e B, 6/04
CX P S o ftw a re V e r sio n 2.0 .
A dditio ns , c h a ng e s o r de le tio ns w e re m a de a t th e s e lo c a tio ns :
1-3, 1-5, 1-31, 1-32, 2-1, 2-8 , 2-9, 2-11, 3-1, 3-6, 3-11, 3-13, 4-4, 4-5, 4-7 , 4-9, 4-10 , 4-12,
5-7 , 5-11, 5-15 & 6-10 .
This document applies to the latest software listed and higher versions. When a subsequent software version
changes the information in this document, a new issue will be released.

PN 624923B
iv
REVISION STATUS

PN 624923B v
CONTENTS
WAR N IN G S AN D PR E CAU TIO N S
R E V ISIO N STATU S, iii
IN TR O D U CTIO N , x iii
U S IN G Y O U R Cy to m ic s F C 50 0 MA N U A LS , x iii
A B O U T T HIS MA N U A L, x iii
CO N V E N T IO N S , x iv
T o Ch o o s e A Co m m a nd W ith T h e K e y b o a rd, x v
D ia lo g B o x , x v
D e s c r ip tio n o f R e p o rting U nits , x v
G R A P HICS , x v
1 SY STE M O V E R V IE W, 1-1
1.1 IN T E N D E D U S E , 1-1
A p p lic a tio ns fo r th e Instr u m e nt, 1-2
1.2 CY T O ME T E R CO N T R O LS A N D IN D ICA T O R S , 1-3
F C 50 0 F lo w Cy to m e try S y s te m , 1-3
Cy to m e te r Indic a to r P a ne l, 1-4
Cy to m e te r R e a dy a nd La s e r O n Indic a to r s , 1-4
S a m p le F lo w R a te Indic a to rs , 1-5
S ig na l A m p litu de Indic a to r s , 1-5
Le v e l S e ns e a nd F lo w Indic a to r s , 1-6
MCL ( Mu lti-tu b e Ca ro u s e l Lo a de r) , 1-6
W o rk s ta tio n, 1-7
P o w e r S u p p ly, 1-7
P r inte r ( O p tio na l) , 1-7
1.3 MICR O S O F T ® W IN D O W S ® D E S K T O P , 1-8
W indo w s 20 0 0 A dm inis tr a to r P a s s w o rd, 1-8
T a s k b a r, 1-8
S ta rt B u tto n, 1-8
R e c y c le B in, 1-9
My Co m p u te r, 1-10
Co ntro l P a ne l, 1-10
U s ing a Mo u s e , 1-10
Co lo r R e s o lu tio n, 1-11
1.4 LE A R N IN G T HE B A S IC O P E R A T IN G T E CHN IQ U E S , 1-13
MCL Ca r o u s e ls , 1-13
B a r-Co de La b e ls , 1-13
P u tting a B a r-Co de La b e l o n a S a m p le T u b e , 1-13
P u tting S a m p le T u b e s in a Ca ro u s e l, 1-14
P u tting a Ca ro u se l in th e MCL, 1-14
R e m o v ing a Ca ro u s e l fro m th e MCL, 1-16

PN 624923B
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CONTENTS
1.5 CX P S O F T W A R E S HO R T CU T S , 1-17
K e y b o a rd S h o r tc u ts, 1-17
D ra g A nd D ro p , 1-18
T o o lb a r B u tto ns , 1-19
1.6 CR E A T IN G P R O T O CO LS , 1-20
1.7 CR E A T IN G R E G IO N S , 1-22
Cre a te P o ly g o na l R e g io ns , 1-22
Cre a te R e c ta ng u la r R e g io ns , 1-23
Cre a te Q u a dra nt R e g io ns , 1-24
R e p o s itio ning a Q u a dr a nt R e g io n, 1-25
Cre a te Line a r R e g io ns , 1-26
Cre a te Mu ltip le Line a r R e g io ns , 1-27
1.8 CR E A T IN G G A T E S , 1-28
Cre a te A u to G a te , 1-28
Co nv e r t a P o ly g o na l R e g io n to a n A u to G a te R e g io n, 1-30
A u to m a tic G a te Cre a tio n, 1-30
1.9 CR E A T IN G F LO W P A G E S , 1-31
F lo w P A G E E x a m p le , 1-31
D e le te F lo w P A G E S , 1-32
1.10 CR E A T IN G P A N E LS , 1-32
U s e P lo ts a nd G a te s fro m P re v io u s T e s t, 1-34
U s e P lo ts a nd G a te s fro m P ro to c o l, 1-34
U s e Ins tru m e nt S e tting s , 1-34
U s e R e g io ns fro m P ro to c o l, 1-34
Ca rry R e g io ns fro m P re v io u s P ro to c o l, 1-34
U s e R e g io ns fro m T h is P ro to c o l:, 1-34
U s e Ins tru m e nt S e tting s fro m P re v io u s T e s t, 1-34
U s e Ins tru m e nt S e tting s fro m E x te r na l F ile , 1-34
P a ra m e te r N a m e s , 1-34
1.11 CR E A T IN G W O R K LIS T S , 1-35
W o rk lis t P a ne l, 1-35
W o r k lis t T e s t, 1-36
1.12 U S IN G T HE O N LIN E HE LP S Y S T E M, 1-37
A c c e s s O nline He lp , 1-37
Ho w to U s e He lp , 1-37
He lp Me nu , 1-38
CX P He lp , 1-38
A b o u t... , 1-38
O nline S u p p o rt, 1-38
1.13 P E R F O R MA N CE CHA R A CT E R IS T ICS , 1-38
2 D AIL Y R O U TIN E , 2-1
2.1 B E F O R E Y O U B E G IN , 2-1

PN 624923B vii
CONTENTS
2.2 D A ILY S T A R T U P, 2-1
Ch e c k W a s te a nd R e a g e nt Le v e ls , 2-2
P o w e r th e Co m p u te r a nd Cy to m e te r O N , 2-3
T u r ning O n P o w e r, 2-3
Lo g g ing O nto W indo w s S o ftw a re , 2-4
Lo g g ing O nto CX P S o ftw a re , 2-4
Ch e c k th e P o w e r S u p p ly, 2-6
A dditio na l S ta rt U p Ch e c k s , 2-8
2.3 D A ILY S HU T D O W N , 2-9
W h e n to S h u t D o w n th e Cy to m e te r, 2-9
B e fo re P e rfo rm ing S h u t D o w n, 2-9
P o w e r th e Co m p u te r a nd Cy to m e te r O F F, 2-9
A fte r Ins tru m e nt S h u t D o w n, 2-11
2.4 E X T E N D E D S HU T D O W N , 2-12
2.5 CY T O ME T E R A U T O S T A R T U P, 2-14
3 Q U AL ITY CO N TR O L , 3-1
3.1 IN T R O D U CT IO N , 3-1
3.2 Q C P R O CE S S E S , 3-1
3.3 D A ILY Q C, 3-2
P re p a r ing A u to S e tu p S a m p le s , 3-2
3.4 A U T O S E T U P S CHE D U LE R , 3-4
3.5 A U T O S E T U P II W IZ A R D , 3-5
B e fo re R u nning th e A u to S e tu p II W iz a rd, 3-5
3.6 A P P LICA T IO N D E F IN IT IO N W IZ A R D , 3-10
B e fo re R u nning th e A p p lic a tio n D e finitio n W iz a rd, 3-10
S e tting U p Q C P ro du c ts , 3-14
E x p o rting R e g io n R e s u lts T o Q C D a ta b a s e a nd A s s ig ning Q C P ro du c ts , 3-14
R e v ie w Q C R e s u lts , 3-15
4 R U N N IN G SAM PL E S, 4-1
4.1 S A MP LE R E Q U IR E ME N T S , 4-1
4.2 B E F O R E R U N N IN G S A MP LE S , 4-1
4.3 R U N N IN G S A MP LE S - MCL A U T O MA T IC MO D E , 4-2
4.4 R U N N IN G S A MP LE S - MCL MA N U A L MO D E , 4-5
4.5 R U N N IN G S A MP LE S - S IN G LE T U B E MO D E , 4-9
5 O PE R ATIO N PR IN CIPL E S , 5-1
5.1 W HA T T HIS CHA P T E R E X P LA IN S , 5-1

PN 624923B
viii
CONTENTS
5.2 S A MP LE F LO W , 5-1
S a m p le Lo a ding , 5-1
Hy dro dy na m ic F o c u s ing , 5-1
5.3 LA S E R B E A M S HA P IN G , 5-3
5.4 CE LL ILLU MIN A T IO N , 5-4
F o r w a rd S c a tte r, 5-4
S ide S c a tte r a nd F lu o re s c e nt Lig h t, 5-4
5.5 LIG HT CO LLE CT IO N , S E P A R A T IO N A N D ME A S U R E ME N T , 5-4
F o r w a rd S c a tte r Co lle c tio n, 5-4
S ide S c a tte r a nd F lu o re s c e nt Lig h t Co lle c tio n, 5-6
S ide S c a tte r, 5-6
F lu o re s c e nt Lig h t, 5-7
5.6 S IG N A L P R O CE S S IN G , 5-8
V o lta g e P u ls e S ig na ls , 5-8
P e a k S ig na l, 5-8
Inte g ra l S ig na l, 5-10
A m p lific a tio n, 5-10
S ig na ls G e ne ra te d, 5-10
5.7 P R O T O CO LS , 5-11
S p e c ia l P ro to c o ls a nd P a ne ls , 5-11
S p e c ia l P a ne ls , 5-11
5.8 A U T O MA T E D S O F T W A R E F E A T U R E S , 5-12
5.9 P A R A ME T E R S , 5-12
A U X ilia ry P a ra m e te r, 5-12
W h e n to U s e th e A U X P a r a m e te r, 5-12
T IME P a ra m e te r, 5-12
R A T IO P a ra m e te r, 5-13
5.10 P LO T D IS P LA Y , 5-13
P R IS M, 5-13
R e g io ns , 5-15
G a ting , 5-15
D a ta S to ra g e , 5-15
His to g r a m S ta tis tic s , 5-16
Line a r R e g io n S ta tis tic s , 5-16
Lo g R e g io n S ta tis tic s , 5-17
Lo g -Lo g Me a n Me th o d, 5-17
Lin-Lo g Me a n Me th o d, 5-18
6 TR O U BL E SH O O TIN G , 6-1
6.1 P R E CA U T IO N S /HA Z A R D S , 6-1
La s e r/R a dia tio n P re c a u tio ns , 6-1
La s e r W a rning La b e ls , 6-1
W a r ning La b e ls o n U P S , 6-10

PN 624923B ix
CONTENTS
D is p o s a l P re c a u tio n, 6-11
6.2 MA IN T E N A N CE S CHE D U LE S , 6-11
Cle a ning S c h e du le , 6-11
R e p la c e m e nt S c h e du le , 6-11
6.3 E R R O R ME S S A G E S , 6-11
D is p la y Lo c a tio ns , 6-11
CX P S o ftw a re S c re e n, 6-11
Cy to m e te r S ta tu s Me s s a g e s S c re e n, 6-12
Cy to m e te r.lo g F ile , 6-12
6.4 E R R O R ME S S A G E CY T O ME T E R .LO G F ILE , 6-13
Ho w to A c c e s s th e Cy to m e te r.lo g F ile , 6-13
F ro m CX P S o ftw a re , 6-13
F ro m W indo w s D e s k to p , 6-13
Cy to m e te r.Lo g E ntr y D e sc rip tio n, 6-14
Ho w to S e a rc h th e Cy to m e te r.lo g F ile , 6-14
O th e r F u nc tio ns A v a ila b le , 6-14
P rint, 6-14
Ch a ng e F o nt, 6-14
F ind He lp T o p ic s , 6-14
Cy to m e te ra rc h iv e .lo g F ile , 6-14
6.5 E R R O R ME S S A G E S T A B LE , 6-15
6.6 LE V E L S E N S E IN D ICA T O R S , 6-20
S h e a th Lo w , 6-20
W a s te F u ll, 6-20
6.7 CY T O ME T E R CO N T R O L W IN D O W CA N N O T B E V IE W E D , 6-20
A H O W TO ..., A-1
A .1 S O F T W A R E P R O CE D U R E S , A -1
T h e s e p ro c e du re s a re in th e S y s te m O v e r v ie w c h a p te r in th e Ins tru c tio ns F o r U s e Ma nu a l., A -1
T h e s e p ro c e du re s a re in th e Q u a lity Co ntro l c h a p te r in th e Ins tru c tio ns F o r U s e Ma nu a l., A -1
T h e s e p ro c e du re s a re in th e U s ing CX P S o ftw a re c h a p te r in th e R e fe re nc e Ma nu a l., A -1
A .2 HA R D W A R E P R O CE D U R E S , A -1
T h e s e p ro c e du re s a re in th e S p e c ia l P ro c e du re s Ma nu a l., A -1
B BAR -CO D E SPE CIFICATIO N S, B-1
B .1 B A R -CO D E S A MP LE ID E N T IF ICA T IO N , B -1
B .2 CO R R E CT P LA CE ME N T O F T HE B A R -CO D E LA B E L, B -1
B .3 B A R -CO D E LA B E L S P E CIF ICA T IO N S , B -2
La b e l S iz e a nd T h ic k ne s s , B -2
S y m b o l D im e ns io ns , B -3
La b e l a nd P rint Q u a lity, B -3

PN 624923B
x
CONTENTS
B .4 B A R -CO D E E R R O R R A T E , B -4
B .5 B A R -CO D E S Y MB O LO G IE S , B -5
B .6 B A R -CO D E LA B LE S , B -6
B a r-Co de La b e l O p tic a l Ch a ra c te r is tic s a t 67 0 nm ± 10 % , B -6
B .7 MCL B A R -CO D E R E A D E R , B -6
B .8 B A R -CO D E D E CO D E R , B -7
B .9 CHE CK S U M A LG O R IT HM, B -7
R E FE R E N CE S, R E FE R E N CE S-1
IN D E X, IN D E X-1
TR AD E M AR K S

PN 624923B xi
ILLUSTRATIONS
5.1 F lo w Ce ll ( Hy dro dy na m ic F o c u s ing ) , 5-2
5.2 La s e r B e a m S h a p ing , 5-3
5.3 O p tic a l S y s te m w ith O p tio na l R e d S o lid-s ta te La s e r, 5-5
5.4 O p tic a l S y s te m w ith O p tio na l He N e La s e r , 5-5
5.5 S ing le La s e r F ilte r B lo c k Co nfig u ra tio n, 5-6
5.6 D u a l La s e r F ilte r B lo c k Co nfig u r a tio n, 5-7
5.7 V o lta g e P u ls e F o rm a tio n, P e a k S ig na l, 5-9
5.8 Inte g r a l a nd P e a k P u ls e s , 5-10
5.9 P ris m P lo t, 5-14
6.1 La s e r La b e ls o n th e S e ns ing Co m p a rtm e nt Co v e r, 6-2
6.2 La s e r La b e ls in th e O p tic a l A re a , F ro nt V ie w , 6-3
6.3 La s e r La b e ls in th e O p tic a l A re a , Inte r io r V ie w , 6-4
6.4 La s e r La b e ls o n th e A rg o n La s e r He a d, 6-5
6.5 La s e r La b e ls fo r O p tio na l R e d S o lid-s ta te La s e r, 6-5
6.6 Lo c a tio n o f th e O p tio na l R e d S o lid-s ta te La se r in th e Ins tru m e nt, R e a r V ie w , 6-6
6.7 La s e r La b e ls fo r O p tio na l R e d He liu m -N e o n La s e r, 6-6
6.8 Lo c a tio n o f th e O p tio na l R e d He liu m -N e o n La s e r in th e Instr u m e nt, R e a r V ie w , 6-7
6.9 La s e r La b e ls o n th e Cy to m e te r B a c k P a ne l, 6-7
6.10 La s e r La b e ls o n th e MCL P ro b e Ho u s ing Co v e r, 6-8
6.11 La s e r La b e ls o n th e MCL B a r-Co de R e a de r, 6-9
6.12 Inte rna tio na l W a r ning S y m b o l Lo c a tio ns , 6-10
6.13 E r ro r Me s s a g e o n CX P S o ftw a re S c re e n, E x a m p le , 6-12
6.14 Cy to m e te r S ta tu s Me s s a g e s W indo w , E x a m p le , 6-12
6.15 E x a m p le o f E r ro r Me ss a g e s in Cy to m e te r.lo g F ile , 6-13
6.16 S h e a th Lo w Indic a to r s , 6-20
6.17 W a s te F u ll Indic a to r s , 6-20
B .1 B a r-Co de La b e l, B -1
B .2 B a r-Co de La b e l P la c e m e nt, B -2
B .3 B a r-Co de La b e l S p e c ific a tio ns , B -3

PN 624923B
xii
TABLES
1.1 A p p lic a tio ns fo r th e Ins tr u m e nt, 1-2
6.1 Cle a ning S c h e du le , 6-11
6.2 E r ro r Me s s a g e s , 6-15
B .1 B a r-Co de S y m b o lo g ie s , B -5
B .2 Co de -R e la te d S p e c ific a tio ns , B -6

PN 624923B xiii
INTRODUCTION
T h is intro du c to ry s e c tio n c o nta ins th e fo llo w ing to p ic s :
r U S IN G Y O U R Cy to m ic s F C 50 0 MA N U A LS
r A B O U T T HIS MA N U A L
r CO N V E N T IO N S , a nd
r G R A P HICS .
USING Y OUR Cy tom ics FC 50 0 MANUALS
T h e m a nu a ls lis te d b e lo w a re a v a ila b le a s P D F file s in th e F C 50 0 CX P s o ftw a re a nd th e
O p e r a to r ’s Ma nu a ls CD -R O M. P rinte d v e r s io ns o f th e s e m a nu a ls a re a ls o a v a ila b le b y o rde r.
U s e th e R eferen ce m a nu a l fo r ins tru m e nt s p e c ific a tio ns a nd info r m a tio n o n ins ta lla tio n a nd
s y s te m o p tio ns .
U s e th e In stru ction s For U se m a nu a l fo r th e da y -to -da y r u nning o f y o u r ins tru m e nt. G o
th ro u g h th e de ta ile d s te p -b y -s te p p ro c e du re s o f s ta rtu p , q u a lity c o ntro l ( Q C) , r u nning
s a m p le s , a na ly z ing da ta , p r inting re p o r ts , re v ie w ing Q C da ta , s h u tdo w n. It c o nta ins s a fe ty
a nd
tro u b le s h o o ting info rm a tio n, e r ro r m e s s a g e s , a s w e ll a s in-de p th info rm a tio n o n th e
p r inc ip le s o f flo w c y to m e try, info r m a tio n a b o u t w h a t y o u r ins tru m e nt do e s , a nd th e m e th o ds
it u s e s .
U s e th e Sp ecial Proced u res m a nu a l to c le a n, re p la c e , o r a dju s t a c o m p o ne nt o f th e ins tru m e nt.
U s e th e G ettin g Started m a nu a l a s a b rie f intro du c tio n to th e s y s te m .
U s e th e M aster In d ex to e a sily lo c a te a to p ic in a ny o f y o u r m a nu a ls .
ABOUT THIS MANUAL
T h e info rm a tio n in y o u r F C 50 0 Ins tr u c tio ns F o r U s e m a nu a l is o rg a niz e d a s fo llo w s :
r Ch a p te r 1 S Y S T E M O V E R V IE W
P ro v ide s th e inte nde d u s e o f th e ins tru m e nt, th e c o ntro ls a nd indic a to r s , a nd
info r m a tio n o n u s ing th e s y s te m ’s s o ftw a r e a nd W o r k s ta tio n.
r Ch a p te r 2 D A ILY R O U T IN E
P ro v ide s info r m a tio n o n do ing da ily p ro c e du re s , s u c h a s S ta rtu p a nd S h u tdo w n.
r Ch a p te r 3 Q U A LIT Y CO N T R O L
P ro v ide s info r m a tio n o n h o w to r u n q u a lity c o ntro l m a te r ia l to v e r ify ins tru m e nt s e tu p .
r Ch a p te r 4 R U N N IN G S A MP LE S
P ro v ide s info rm a tio n o n h o w to r u n p a tie nt b lo o d sa m p le s .
s Ch a p te r 5, O P E R A T IO N P R IN CIP LE S
Co nta ins a de s c rip tio n o f flo w c y to m e tr y, th e no r m a l s a m p le flo w th ro u g h th e
ins tr u m e nt, h o w lig h t c o lle c tio n a nd sig na l p ro c e ss ing a re a c c o m p lis h e d a nd h o w th e
p a ra m e te r s a re de riv e d.
r Ch a p te r 6, T R O U B LE S HO O T IN G
D e s c r ib e s la s e r s a fe ty p re c a u tio ns a nd th e lo c a tio n o f th e la s e r-re la te d la b e ls a nd h o w to
tro u b le s h o o t th e s y ste m w ith th e e rro r m e s s a g e s th a t a p p e a r o n th e s c re e n.

PN 624923B
xiv
INTRODUCTION
CONVENTIONS
s A p p e ndic e s
T h e a p p e ndic e s p ro v ide re fe re nc e m a te ria l o n th e fo llo w ing to p ic s :
r HO W T O ...
r B A R -CO D E S P E CIF ICA T IO N S
r IN D E X
P ro v ide s p a g e nu m b e r s fo r inde x e d info rm a tio n.
CONVENTIONS
T h is m a nu a l u s e s th e fo llo w ing c o nv e ntio ns :
r T h ro u g h o u t th is m a nu a l y o u r F C 50 0 is a ls o re fe r re d to a s th e s y s te m o r ins tru m e nt.
r Bold font indic a te s a s o ftw a re o p tio n, s u c h a s Cy tom eter.
r Italics font indic a te s sc re e n te x t dis p la y e d o n th e ins tru m e nt, s u c h a s
P reparing S amples.
r Courier font indic a te s te x t y o u h a v e to ty p e u s ing th e k e y b o a rd.
r indic a te s a k e y ( s u c h a s Û) .
r + indic a te s th a t th e tw o k e y s lis te d (s u c h a s Þ+Ê) a r e link e d fo r a s p e c ific
fu nc tio n a nd m u s t b e p re s s e d in th is s e q u e nc e :
a . P re s s do w n o n th e firs t k e y lis te d a nd w h ile c o ntinu ing to p re s s it, p re s s do w n o n
th e s e c o nd k e y lis te d.
b . R e le a s e b o th k e y s a t th e s a m e tim e .
r indic a te s to p re s s a nd re le a s e th e fir s t k e y lis te d th e n p re s s a nd re le a s e th e ne x t
k e y lis te d. F o r e x a m p le : YÛ.
r Ic o ns /b u tto ns to s e le c t fu nc tio ns o n th e s o ftw a re s c re e n a re s h o w n w ith in te x t.
E x a m p le : .
r indic a te s to u s e th e m o u s e to s e le c t th e s c re e n b u tto n la b e le d
. S e e U s ing a Mo u s e .
r File ttSa v e indic a te s to u s e th e m o u s e to s e le c t th e Sa v e ite m o n th e File m e nu .
r Éth ro u g h Ô a re s p e c ia l fu nc tio n k e y s.
r A N ote c o nta ins info rm a tio n th a t is im p o rta nt to re m e m b e r o r h e lp fu l in p e r fo r m ing a
p ro c e du re .
r T h e te rm s “ s c re e n” a nd “ w indo w ” a re u s e d inte rc h a ng e a b ly.
r
SHOW ME
m e a ns th e re is a v ide o a v a ila b le fo r th e p ro c e du re in th e o nline h e lp .

PN 624923B xv
INTRODUCTION
G R A P H ICS
To Ch oose A Com m a nd With Th e Key b oa rd
A fte r y o u p re s s Þ, e a c h m e nu na m e h a s o ne le tte r u nde r line d to indic a te w h ic h le tte r to u s e
to p u ll do w n th e m e nu . F o r e x a m p le , th e le tte r F in th e F ile m e nu is u nde r line d, p re s s Fto
p u ll do w n th e F ile m e nu ; th e le tte r E in th e E dit m e nu is u nde r line d, p re s s Eto p u ll do w n
th e E dit m e nu .
Dia log Box
D ia lo g b o x e s re c e iv e c o m m a nds o r info r m a tio n; fo r e x a m p le , a file na m e dia lo g b o x re c e iv e s
info rm a tio n a b o u t a file na m e .
Descrip tion of Rep orting Units
U nle s s o th e r w is e s ta te d, a ll p a r a m e te r u nits a re s h o w n in th e U S u nit fo r m a t ( c e lls /µ L)
th ro u g h o u t th e m a nu a ls .
GRAPHICS
A ll g r a p h ic s , inc lu ding s c re e ns a nd p rinto u ts , a re fo r illu s tra tio n p u rp o s e s o nly a nd m u s t no t
b e u s e d fo r a ny o th e r p u rp o s e .
Comman d Fu n ction
ÛA c c e p ts y o u r s e le c tio n.
ÈS to p s th e o p e r a tio n, dis c a rding y o u r c h o ic e s .
ÙMo v e s c u r s o r o v e r diffe re nt c h o ic e s if th e re a re m u ltip le o p tio ns - s e e
W indo w s ®m a nu a ls fo r W indo w s o p e r a tio n v ia k e y b o a rd.
Þ+Ù W h e n y o u h a v e m o re th a n o ne a p p lic a tio n W indo w o p e n, u s e Þ+Ù to
s w itc h b e tw e e n ta s k s .
A c c e p ts th e info r m a tio n y o u h a v e s e le c te d o r ty p e d.
S to p s th e o p e ra tio n, ig no r ing y o u r c h o ic e s .

PN 624923B
xvi
INTRODUCTION
G R A P H ICS

PN 624923B 1 -1
1
1SY STEM OVERVIEW
T h e info r m a tio n in th is s e c tio n inc lu de s :
r IN T E N D E D U S E
r CY T O ME T E R CO N T R O LS A N D IN D ICA T O R S
r MICR O S O F T ® W IN D O W S ® D E S K T O P
r LE A R N IN G T HE B A S IC O P E R A T IN G T E CHN IQ U E S
r CX P S O F T W A R E S HO R T CU T S
r CR E A T IN G P R O T O CO LS
r CR E A T IN G R E G IO N S
r CR E A T IN G G A T E S
r CR E A T IN G F LO W P A G E S
r CR E A T IN G P A N E LS
r CR E A T IN G W O R K LIS T S
r U S IN G T HE O N LIN E HE LP S Y S T E M
r P E R F O R MA N CE CHA R A CT E R IS T ICS
1.1 INTENDED USE
T h e Cy to m ic s F C 50 0 is a s y s te m fo r th e q u a lita tiv e a nd q u a ntita tiv e m e a s u r e m e nt o f
b io lo g ic a l a nd p h y s ic a l p ro p e rtie s o f c e lls a nd o th e r p a rtic le s . T h e s e p ro p e rtie s a re m e a s u re d
w h e n th e c e lls p a s s th ro u g h o ne o r tw o la s e r b e a m s in s ing le -file .
T h e ins tr u m e nt c a n sim u lta ne o u s ly m e a s u re fo r w a rd sc a tte r, s ide s c a tte r, a nd fiv e flu o re s c e nt
dy e s u s ing o ne o r tw o la s e r s a t 48 8 nm a nd e ith e r 635 nm ( S o lid-s ta te la s e r) o r 633 nm
( He N e la s e r ) . T h e re fo re , th e ins tru m e nt c a n p e rfo r m c o r re la te d m u ltip a r a m e te r a na ly s e s o f
indiv idu a l c e lls .
IMPORTANT T h e us e of th e d ata g enerated b y th is ins trument d ep end s up on th e reg ulatory s tatus of th e
reag ents y ou us e. If th e reag ent is lab eled b y th e manufacturer “ For R es earch U s e O nly . Not for us e in
d iag nos tic p roced ures ,” U S fed eral law p roh ib its th e us e of th e d ata for d iag nos is .
If, h ow ever, th e reag ent s ub s y s tem is lab eled “ For in vitro d iag nos tic us e,” th e entire s y s tem can b e
cons id ered a med ical d evice for g enerating d ata to b e us ed for d iag nos is .

PN 624923B
1 -2
SY STEM OVERVIEW
INTEND ED U SE
Ap p lica tions for th e Instrum ent
T h e F C 50 0 is c o nside re d a n e q u iv a le nt s y s te m to th e CO U LT E R ® E P ICS ® X L™ a nd th e
CO U LT E R E P ICS X L-MCL™ flo w c y to m e te r s w h e n u s e d w ith th e fo rw a rd sc a tte r F ie ld S to p
in th e fa c to r y s e t p o s itio n ( 1-19°) .
T a b le 1.1 lis ts th e m a ny a p p lic a tio ns fo r th e ins tru m e nt. In a dditio n to h u m a n c e lls , o th e r c e ll
ty p e s c a n b e a na ly z e d, s u c h a s :
r P la nt c e lls
r Ma r ine p la nk to n
r A nim a l c e lls
r B a c te ria
Ta b le 1.1 Ap p lica tions for th e Instrum ent
Ap p lica tions Cell Ty p es
Sa m p le Prep a ra tion
(refer to th e rea g ent's
p a ck a g e insert) Mea surem ents Pa ra m eters
Cell s urface
antig ens
W h ole b lood
Buffy coats
M ononuclear cells
D is s ociated tis s ue
Platelets
Bone M arrow
ImmunoPrep ™reag ents w ith
th e Q -PR E P™W orks tation,
th e M ulti-Q -Prep ™
W orks tation or th e T Q -Prep ™
W orks tation
W h ole b lood ly s ing reag ent
kit
Cell g rad ient s ep aration
Fluores cent-lab eled
antib od ies
T h romb o Fix™ p latelet
s tab iliz er
Cell s iz e and
g ranularity
FIT C, R D 1 , E CD , PC5,
PC7 , APC and
APC-CY 7 , PC7
Forw ard and s id e
s catter (L og /L inear)
Fluores cence one,
tw o, th ree, four and
five (L og /L inear)
Pris m
R atio
T ime
AU X /Peak
Nucleic acid s W h ole b lood
D is s ociated tis s ue
Froz en s ections
Paraffin s ections
Bod y fluid s
V arious s taining meth od s ,
includ ing :
E th id ium b romid e
Prop id ium iod id e
D NA Prep reag ents
L eukaS ure™
Cell s iz e and
g ranularity
G reen, red , and orang e
fluores cence
Forw ard and s id e
s catter
Fluores cence one,
tw o, th ree, four and
five (L og /L inear)
R atio, AU X Peak
K inetics W h ole b lood
Buffy coats
M ononuclear cells
T is s ue culture cells
V arious meth od s , includ ing :
Fluo 3 fluores cence
Cell s iz e and
g ranularity
G reen fluores cence
T ime
Forw ard and s id e
s catter
Fluores cence one
R atio
T ime

PN 624923B 1 -3
SY STEM OVERVIEW
CY TOM ETER CONTR OL S A ND IND ICA TOR S 1
1.2 CY TOMETER CONTROLS AND INDICATORS
FC 50 0 Flow Cy tom etry Sy stem
CH
Cell function W h ole b lood
Buffy coats
M ononuclear cells
T is s ue culture cells
V arious meth od s and d y es ,
includ ing :
D CFH -D A
D iO C5(3)
FD A
Cell s iz e and
g ranularity
G reen fluores cence
Forw ard and s id e
s catter
Fluores cence one
R eticulocy tes W h ole b lood T h iaz ole orang e
corip h os p h ine O (CPO )
Acrid ine O rang e
Cell s iz e and
g ranularity
G reen fluores cence
R ed fluores cence
L og /L inear Forw ard
and S id e S catter
L og Fluores cence one
L og Fluores cence four
Ta b le 1.1 Ap p lica tions for th e Instrum ent
(Continued)
b
c
d
e
f
g
h
i
j
557 1 026B

PN 624923B
1 -4
SY STEM OVERVIEW
CY TOM ETER CONTR OL S A ND IND ICA TOR S
Cy tom eter Ind ica tor Pa nel
Cy tom eter Rea d y a nd La ser On Ind ica tors
bMCL gMo u s e
cCy to m e te r hU S B b o x
dHe N e la s e r o p tio n iCo m p u te r
eK e y b o a rd jP o w e r S u p p ly
fMo nito r
bB lu e m e a ns th e A rg o n la s e r is o n.
cR e d m e a ns th e re d la s e r s h u tte r is o p e n.
dG re e n m e a ns Cy to m e te r is o n a nd in th e R e a dy s ta te .
d c b